Persistent pain is a highly prevalent, global cause of disability. Research suggests that many healthcare professionals are not well equipped to manage pain and that this may be attributable at least in part to undergraduate education. The primary aim of this study was to quantify and compare first and final year nursing, midwifery and allied health professional (NMAHP) students' pain-related knowledge and attitudes. The secondary aim was to explore the factors influencing students' pain-related knowledge and attitudes. This cross-sectional study included 1154 first and final year healthcare students, from 12 universities in five different countries. Participants completed the Revised Neurophysiology of Pain Quiz (RNPQ) [knowledge] and the Health Care Providers Pain and Impairment Relationship Scale (HC-PAIRS) [attitudes]. Physiotherapy was the only student group with statistically and clinically improved pain-related knowledge [mean difference, 95% CI] (3.4, 3.0 to 3.9, p=0.01) and attitudes (-17.2, -19.2 to 15.2, p=0.01) between first and final year. Pain education teaching varied considerably from course to course (0 to 40 hours), with greater levels of pain-related knowledge and attitudes associated with higher volumes of pain-specific teaching. There was little difference in pain knowledge and attitudes between all first and final year NMAHP students other than physiotherapy. This suggests that for most NMAHP disciplines, undergraduate teaching has little or no impact on students' understanding of pain. There is an urgent need to enhance pain education provision at the undergraduate level in NMAHPs.